Methodology
sample preparation
9
How to place the solution into a petri dish!
500mL
300mL
100mL
16 LB
16 LB/amp
8 LB/amp/ara
Starting with the 500mL of LB agar solution, made on page 8, pour 12.5mL into 16 individual petri dishes.
In the remaining 300mL, add the rehydrated ampicillin, and pour 12.5mL of the solution into another 16 petri dishes.
With the remaining 100mL, mix in the rehydrated arabinose. Distrubute the remaining solution among 8 more petri dishes; 12.5mL in each.
to grow a starter plate before begining the experiment on the next page!
DON'T
FORGET
To do this, rehydrate lyophilized E.coli by adding 0.25mL of transformation solution directly into the vial, using a sterile pipette. Recap the vial and allow it to sit for 5 minutes. Shake to mix, then streak 1 LB agar plate with the rehydrated E.coli, using a sterile inoculation loop.
Make sure to cover as much surface area on the agar, as possible.
Place the covered plate upside down in an incubator at 37C, for 24-36 hours, before use.
Each cell on the plate will multiply to become millions of genetically identical cells.
NOTE: these measurements are enough for multiple experiments to be preformed.
1 experiment will require:
2 LB plates, 2 LB/amp plates and 1 LB/amp/ara plate.
Final concentrations per plate:
6mg/mL of arabinose
0.1mg/mL of ampicillin